| Objective:The mouse skin aging animal model was established by subcutaneous injection of D-galactose combined with Ultraviolet radiation,and the effects of microne combined with rehabilitation solution on skin inflammatory factors and Langerhans cells in aging mice were preliminaries explored.Method:Fifty healthy male mice were randomly divided into 5 groups: blank control group（A）,aging model group（B）,microneedle combined with kangfuxin solution group（C）,microneedle combined with kangfuxin solution group（D）,and kangfuxin solution group（E）,with 10 mice in each group.Except for group A,mice in B、C、D、E four groups were subcutaneously injected 5% D-galactose（10m L /kg）into the neck and back skin daily,combined with UV（UVA+UVB）irradiation for 5 days A week,until the end of the experiment.From day 11 of the experiment,group B: no special intervention except subcutaneous injection of D-galactose and UV irradiation.Group C: 0.5mm microneedles were rolled on the back of the nose of the mice,and kangfuxin liquid was sprayed on the treatment site,kept moist for 20 minutes,once every 14 days,4 times in total,and sprayed externally once every day during the interval,at the same time as before;Group D: after microneedle treatment,normal saline was sprayed externally for 20 minutes,once every 14 days,4 times in total.During the interval,normal saline was sprayed externally once a day,at the same time as before.Group E: Separate external spray of rehabilitation solution,keep moist for20 minutes,once a day.After the experiment,the skin appearance changes of mice were observed,hematoxylin-Eosin（HE）staining and Masson staining were observed to observe the epidermal and dermal tissue structure changes,AND the contents of interleukin-1β（IL-1β）、interleukin-6（IL-6）and Tumor necrosis factor-α（TNF-α）were detected by Enzyme-linked immunoadsorption assay（ELISA）.The number and morphology of Langerhans cells（LC）were observed by immunofluorescence method.Result:（1）Naked eye observation: except group A,the skin of mice in other groups showed signs of aging.Compared with group A,the skin of mice in group B was rough,thickened,desquamate,skin elasticity disappeared,and the skin photoaging score was the highest（P < 0.05）,suggesting that the ICR mouse skin aging model was successfully constructed.Compared with group B,skin appearance of mice in groups C,D and E was improved,and skin photoaging score was decreased（P < 0.05）.Compared with group D and GROUP E,group C had the most obvious improvement,the skin light aging score was the lowest（P < 0.05）,and the appearance change was closer to group A.（2）HE and Masson staining: the epidermal keratinocytes of mice in group B were hyperplasia,dermal collagen fibers were reduced,loose and denatured,blood vessels in the superficial dermis were dilated and hyperemic,and there were more lymphocytes around accessory organs.Compared with group B,the pathological manifestations of skin aging tissues in groups C,D and E were all reduced,and group C was the most significantly reduced compared with groups D and E.（3）The contents of inflammatory cytokines IL-1β,IL-6 and TNF-α in group B were significantly increased compared with group A（P < 0.05）;Compared with group B,the content of groups C,D and E was significantly decreased（P < 0.05）,and the content of group C was the lowest compared with groups D and E（P < 0.05）.（4）Langerhans cells: Compared with group A,the number of LC in group B was significantly reduced（P < 0.05）,and the morphology was abnormal;Compared with group B,the number of LC in group C,D and E increased（P < 0.05）,and the morphology was roughly normal.Compared with group D and E,the number of LC in group C was the largest（P < 0.05）,and the morphology was basically normal.Conclusion :The combination of microneedle and kangfuxin solution can effectively improve skin aging,and the anti-skin aging mechanism may be related to repairing the skin structure of mouse skin aging model,reducing the content of IL-1β,IL-6 and TNF-α,increasing the amount of LC and repairing morphology. |
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